Bacterial co-infection and early mortality among pulmonary tuberculosis patients in Manila, The Philippines.

OBJECTIVE: To investigate the prevalence of bacterial co-infection and its effect on early mortality among hospitalised human immunodeficiency virus (HIV) negative pulmonary tuberculosis (PTB) patients in Manila, the Philippines. DESIGN: A prospective observational study was conducted at a national infectious disease hospital. HIV-negative PTB patients aged 13 years hospitalised from November to December 2011 and from December 2012 to May 2013 were enrolled. Sputum samples were tested for Mycobacterium tuberculosis and six respiratory bacterial pathogens using polymerase chain reaction (PCR). RESULTS: Of 466 patients, 228 (48.9%) were TB-PCR-positive. Overall, bacterial pathogens in purulent sputum were detected in 135 (29.0%) patients: Haemophilus influenzae was the most common bacterium (21.2%), followed by Streptococcus pneumoniae (7.9%). The prevalence of bacterial co-infection did not differ between TB-PCR-positive and -negative patients. A total of 92 (19.7%) patients died within 2 weeks. Bacterial co-infection was significantly associated with an increased risk of 2-week mortality among TB-PCR-positive patients (adjusted risk ratio [aRR] 1.67, 95%CI 1.03-2.72). This association was also observed but did not reach statistical significance among TB-PCR-negative patients (aRR1.7, 95%CI 0.95-3.02). CONCLUSION: Bacterial co-infection is common and contributes to an increased risk of early mortality among HIV-negative PTB patients.

Risk factors for death among hospitalised tuberculosis patients in poor urban areas in Manila, The Philippines.

OBJECTIVE: To determine the mortality rate and risk factors for in-hospital death among hospitalised human immunodeficiency virus (HIV) negative tuberculosis (TB) patients in poor urban areas in the Philippines. DESIGN: A cross-sectional study was conducted at a national infectious disease hospital in Manila City. The target population was patients aged ≥ 13 years with all forms of HIV-negative TB admitted from October to December 2009. Demographic and clinical information was collected from medical charts, and risk of in-hospital death was measured. RESULTS: Of 407 HIV-negative TB patients, four were excluded due to missing records, and 403 were included in the analysis. The majority were poor urban residents (90%), and 66% were males. Overall, 37.5% of hospitalised patients died in the hospital (151/403), 30% of whom died before the third day of hospitalisation. Risk factor analysis demonstrated that complications of bacterial pneumonia had the greatest effect on in-hospital death (aOR 4.53, 95%CI 2.65-7.72), followed by anorexia (aOR 3.01, 95%CI 1.55-5.84), anaemia (haemoglobin <10 g/dl, aOR 2.35, 95%CI 1.34-4.13) and older age (aged ≥ 50 years, aOR 1.85, 95%CI 1.08-3.17). The presence of haemoptysis (aOR 0.44, 95%CI 0.25-0.80) was associated with improved survival. CONCLUSION: Mortality among hospitalised HIV-negative TB patients was extremely high in poor urban areas in the Philippines.

Thalidomide attenuates learning and memory deficits induced by intracerebroventricular administration of streptozotocin in rats.

Neuroinflammatory responses caused by amyloid ß (Aß) peptide deposits are involved in the pathogenesis of Alzheimer's disease (AD). Thalidomide has a significant anti-inflammatory effect by inhibiting TNF-α, which plays role in Aß neurotoxicity. We investigated the effect of thalidomide on AD-like cognitive deficits caused by intracerebroventricular injection of streptozotocin (STZ). Intraperitoneal thalidomide was administered 1 h before the first dose of STZ and continued for 21 days. Learning and memory behavior was evaluated on days 17, 18 and 19, and the rats were sacrificed on day 21 to examine histopathological changes. STZ injection caused a significant decrease in the mean escape latency in passive avoidance and decreased improvement of performance in Morris water maze tests. Histopathological changes were examined using hematoxylin-eosin and Bielschowsky staining. Brain sections of STZ treated rats showed increased neurodegeneration and disturbed linear arrangement of cells in the cortical area compared to controls. Thalidomide treatment attenuated significantly STZ induced cognitive impairment and histopathological changes. Thalidomide appears to provide neuroprotection from the memory deficits and neuronal damage induced by STZ.

Honey as a substitute for formalin?

Formalin has long been the standard fixative for clinical routines worldwide. After the Formaldehyde Standard became law in the US in 1987, as a result of increasing concerns about the potential carcinogenicity of formaldehyde, attempts have been made to find safer alternatives. Alcoholic formalin is a useful fixative, because in addition to fixation, dehydration also is begun. For centuries, honey has been known to be an antibacterial agent with the potential to preserve compounds without harmful effects on its users. We compared the effects of honey fixation with other routine fixatives using conventional histochemical and immunohistochemical staining methods. Our results demonstrated that tissues fixed in either honey or alcoholic formalin and 10% neutral buffered formalin (NBF) have similar histomorphology. Honey fixation showed minor histomorphological differences among the various tissues; however, it did not influence affect correct diagnostic conclusions. Our results suggested that honey can be used as a safe alternative to formalin in histopathology.

Increased in vitro Cell Proliferation by Chitosan/pGM-CSF Complexes.

Granulocyte macrophage colony stimulating factor, a potent hematopoietic cytokine, has been shown to stimulate production of white blood cells following chemotherapy. Therefore, the granulocyte macrophage colony stimulating factor gene is a potential candidate for the treatment of different pathological conditions. The purpose of this study is to investigate the suitability of chitosan as carrier for pORF-hGMCSF plasmid encoding granulocyte macrophage colony stimulating factor gene and also to study the effect of complexes on protein production and cell proliferation. Chitosan/pGM-CSF complexes were prepared using different (+/-) ratios (from 0.01/1 to 5/1). Complex formation was checked with agarose gel electrophoresis. The size and zeta potential values were measured. Enzyme and serum stability of complexes were studied. In vitro transfection properties of complexes were studied in HeLa cells. According to agarose gel electrophoresis, full complexation was obtained at 0.1/1 and higher chitosan/pGM-CSF ratios. Complexes having about 132 nm size and +13.7 mV zeta potential value were obtained. Chitosan complexes protected plasmid against enzymatic and serum effects. The gene expression-dependent cell proliferation after transfection of chitosan/pGM-CSF complexes at 72 h was markedly increased in comparision with the level of control group. These results indicate that the effect of chitosan/pGM-CSF complexes on cell proliferation was changed with N/P ratio and time-dependently. For GM-CSF therapy, chitosan/pGM-CSF complexes may be used as alternative to conventional protein treatments. Chitosan may be a good carrier for pORF-hGMCSF. Further, in vivo study is ongoing.

In vitro silencing effect of chitosan nanoplexes containing siRNA expressing vector targeting VEGF in breast cancer cell lines.

Small interferring RNA (siRNA) is a powerful tool for controlling cellular processes of gene silencing at post-transcriptional level due to its high sequence-specific inhibition efficiency. The aim of this study is to compare silencing effect of chitosan/shVEGF (shRNA-expressing plasmid DNA targeting vascular endothelial growth factor-A (VEGF-A)) nanoplexes between four different cell lines, two of which are MCF-7 and MDA-MB435 breast cancer cell lines. Nanoplexes were prepared using different concentrations. The morphological and physicochemical characterization were made and silencing activity of appropriate formulations were investigated in vitro. As a result of in vitro transfection studies made in different cell lines with nanoplexes in the different ratios, the highest gene inhibition (60%) was measured in MCF-7 after transfection while the lowest gene inhibition (29%) was observed in MDA-MB435. This work suggests that RNA interference has potential to be applied to delivery system studies and to the angiogenesis treatment.